Quantification of mitochondrial DNA copy number: Pre-analytical factors |
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Authors: | Antonio L Andreu Ramiro Martinez Ramon Marti Elena García-Arumí |
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Institution: | aUnitat de Patologia Mitocondrial, Centre d’Investigacions en Bioquímica i Biologia Molecular (CIBBIM), Institut de Recerca Hospital Universitari Vall d’Hebron, Barcelona, Spain;bCentro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), ISCIII, Barcelona, Spain |
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Abstract: | Mitochondrial DNA (mtDNA) content is important for understanding many cellular processes. Several pre-analytical factors, from sample collection to DNA extraction can affect measurement of mtDNA copy number. In the present study, whole blood samples yielded a higher mtDNA copy number than buffy coat samples. mtDNA content is affected by the cell separation method used and the time between blood withdrawal and cell separation. Thus, reference values must be established with the same type of sample. As to the DNA isolation and purification method, the manual phenol method can give randomly false high values. The QIAamp DNA Mini Kit provided the most highly reproducible mtDNA/nDNA yield. |
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Keywords: | mtDNA depletion syndrome Mitochondrial DNA Real-time PCR DNA isolation and purification methods |
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